Re: Quenching Rhodamine

From: Howard Shapiro (hms@shapirolab.com)
Date: Thu Jan 21 1999 - 19:39:52 EST


Scott Menzie asks:
>     
>     Does anyone out there know a good way to quench rhodamine.  Ive got a 
>     researcher doing phagocytosis assays and needs to quench rhodamine 
>     labeled beads which have not been phagocytized. Does trypan blue work? 
>     If it does or anything else does what concentration do you recommend?  
>     Thanks in advance for your help!!
>     

For any quencher to work, the beads would have to be labeled only on the
surface.  The "rhodamine" beads from polysciences, and many other
fluorescent beads, are impregnated with dye, which, being in the polymer
matrix, is not accessible to a water-soluble quencher like trypan blue, and
it would probably not be easy to find an hydrophobic quencher which would
get into the beads from an aqueous solution.  That's why a lot of people who
do phagocytosis assays work with bacteria surface labeled by incubation with
FITC, etc.; the fluorescence of these particles can be quenched.

-Howard



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